D3.1. Whealbi. Wheat and barley Legacy for Breeding Improvement. Grant agreement number: FP Collaborative Project SEVENTH FRAMEWORK PROGRAMME

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1 Whealbi Wheat and barley Legacy for Breeding Improvement Grant agreement number: FP Collaborative Project SEVENTH FRAMEWORK PROGRAMME Deliverable Report on phenotypic evaluation of the basic adaptive traits involved in adaptation to climatic conditions for the whole WHEALBI germplasm Due date: M24 Actual submission date: M26 Project start date: January 1 st, 2014 Duration: 60 months Workpackage concerned: 3 Concerned workpackage leader: CREA Dissemination level: PU 1 / 10

2 Table of contents TABLE OF CONTENTS... 2 GLOSSARY AND DEFINITIONS... 3 DESCRIPTION OF THE DELIVERABLE Objectives Rationale and team involved Main results Conclusions Foreword: The short delay in the submission is due to the decision taken by the WP leader in response to the inputs of the project advisory board of scoring some more traits in addition to those indicated in the DoW. The score of these traits, mainly measured on harvested plants, has slowed down the whole phenotyping process, but it has increased the scientific value of the work. 2 / 10

3 Glossary and Definitions Garden Experiment: a network of nursery field trials carried out in different locations with small plots for the evaluation of basic adaptive traits (e.g. plant height, flowering time). 3 / 10

4 Description of the deliverable The WP3 explores different phenotyping strategies to complement the sequence data generated in WP2. The first phenotyping step has been dedicated to the evaluation of the basic adaptive traits of the whole germplasm collection to identify loci controlling local adaptation in wheat and barley Objective The objective of is to provide a phenotypic evaluation of basic adaptive traits involved in wheat and barley adaptation to different climatic conditions Rationale and teams involved This activity has been accomplished through the organization of a network of field trials (common garden experiments) across different latitudes (from Scotland to Turkey) and climatic conditions (from wet to extreme dry, from sea level to 1200 meters), where all the WHEALBI accessions have been tested. Overall, 10 common garden experiments have been successfully carried out for barley and 8 for wheat. The seeds were distributed from IPK and all trials were sown according to a common augmented prep design prepared by DLO. According to the experimental design, in each location a subset of the accessions (about 20%) was replicated twice. Two elite cultivars were used to control for spatial effects. The garden experiments for wheat were carried out in winter sowing in Hungary (MTAATK), France (INRA), UK (KWS), Turkey (Univ. of Curkurova as subcontractor of IPK) and in spring sowing in Hungary and Turkey. An additional wheat garden experiment was sown in UK during spring, but germination and plant establishment failed due to an exceptionally dry season. The wheat spring sowing in France (INRA) has been converted in the measure of the carbon isotope discrimination (done on winter wheat same location) in response to a specific request of the project advisory board. Furthermore, a wheat trial was carried out also in Israel (HU) to test for adaptation to local conditions and for seed multiplication in view of the activity of Task 3.2, nevertheless this trial did not follow the general experimental design due to rules and limitations imposed by the quarantine requested to import the seeds into Israel as described in details in the first annual report. The garden experiments for barley were carried out using both winter and spring sowing in Hungary (MTAATK), Italy (CREA), UK (JHI) and Turkey (Univ. of Curkurova as subcontractor of IPK). In Turkey all trials were duplicated and carried out in both irrigated and rainfed conditions. Meteo data have been collected in all the environments hosting the common garden experiments Main results In all experiments the plants were scored following common protocols for plant establishment, winter survival, flowering time, plant height, awn length, rowtype (barley). Furthermore, some additional traits (carbon isotope discrimination, seed size, seed weight, diseases, leaf size, peduncle length, fruiting efficiency, etc.) were 4 / 10

5 scored in a limited number of trials. Notably, the additional traits have been included in the WP3 activity in response to specific inputs of the project advisory board and these scores exceed the list of the characters indicated in the DoW. All data have been collected using a standard file format (Ephesys, prepared by INRAURGI) that will facilitate the storage of the data in the WHEALBI website and their analysis. A list of the traits successfully collected in each garden experiment is reported in the following table; for each trait the average value and the range of variation is indicated. Some of the additional traits deriving from the analysis of the plants and seeds harvested from the garden experiments will be completed during the next months (indicated as in progress in the table). Although the phenotyping of some of these traits is still in progress, the data collected so far are sufficient to fulfill the deliverable 3.1 as indicated in the DoW and to complete all the next project activities that relay on the output of the garden experiments. 5 / 10

6 Garden experiment summary. For each crop specie (barley, wheat), sowing time (winter, spring) and location the table reports the traits successfully scored with mean value and range of variation. Barley Winter mandatory traits additional traits BW1 BW2 BW3 BW4 BW5 Variable Name Description CRAITA JHIUK MTAHUN TUR TUR Plant Establishment Visual score (03) Winter survival Visual score (09) Heading date Plant height Nr. Of days from sowing to Zadoks DGS 53 cm from soil to the insertion of Mean=1.3 Max=4; Mean=1.3 Min=156; Max=207; Mean=186.7 Min=27; Max=130; Mean=75.0 Max=2; Mean=1.0 Max=6; Mean=2.9 Min=201; Max=240; Mean=221.2 Min=32; Max=111; Mean=69.9 Mean=2.8 Max=9; Mean=3.1 Min=180; Max=207; Mean=194.6 Min=42; Max=111; Mean=75.2 Mean=1.8 Mean=1.9 Min=169; Max=188; Mean=179.6 Min=60; Max=131.5; Mean=95.1 Min=169; Max=185; Mean=178.8 Min=38.5; Max=128; Mean=89.3 Row type 2rows/6rows/other Available Available Available Available Available Awn length Awn roughness (1000)Grain weight Flag leaf length Flag leaf width Peduncle length Peduncle extrusion Spike length Fruiting efficiency Seed Width Seed Length Seed Area cm from the tip of the spike to the end of the awn 1: Smooth; 2: smooth base/rough tip; 3: rough g average weight of 3 reps of 100 kernels cm from auricles to the tip cm at the widest point cm from the last node to the base of cm from the auricle of the flag leaf to the base of cm from base to tip of Grain/spike weight ratio mm Marvin at JHI, mm Marvin at JHI, mm2 Marvin at JHI, Max=25.0; Mean=12.1 Max=18.5; Mean=10.9 Available Min=30.3 (no H. Spont); Max=69.6; Mean=49.1 Min=3.0; Max=32.0; Mean=13.0 Min=0.2; Max=1.9; Mean=0.7 Min=5.0; Max=65.0; Mean=28.3 Min=1.0; Max=40.0; Mean=5.5 Min=3.0; Max=18.0; Mean=9.9 Min=0.28; Max=0.93; Mean=0.68 Min=23.5; Max=81.3; Mean=52.9 Min=16.5; Max=75.5; Mean=49.5 Min=4.5; Max=37.3; Mean=9.4 Min=3.0; Max=15.5; Mean=8.0 Min=6.0; Max=53.0; Mean=26.1 Min=8.0; Max=69.0; Mean=25.0 Min=2.3; Max=4.4; Mean=3.6 Min=6.3; Max=16.4; Mean=9.9 Min=16.3; Max=39.7; Mean=26.2 Meteo data Available Available Available Available Available Sowing date 27/10/ /10/ /10/ /11/ /11/2014 Notes Bad establ. + insect damage Severe lodging, not harvested Irrigated Rainfed Seeds Stored 10 ears harvested Stored + sent to CREA Stored Stored 6 / 10

7 mandatory traits additional traits Barley Spring BS1 BS2 BS3 BS4 BS5 Variable Name Description CRAITA JHIUK MTAHUN TUR TUR Plant Establishment Heading date Plant height Visual score (03) Nr. Of days from sowing to Zadoks DGS 53 cm from soil to the insertion of Mean=1.5 Available Mean=2.1 Min=67; Max=106; Mean=82.8 Min=47; Max=131; Mean=91.5 Mean=2.9 Min=60; Max=93; Mean=72.5 Min=34; Max=99; Mean=67.9 Max=2; Mean=1.0 Max=2; Mean=1.0 Min=20; Max=93; Mean=52.6 Min=20; Max=113; Mean=66.1 Row type 2rows/6rows/other Available Available Available Awn length (1000)Grain weight Peduncle length Peduncle extrusion Spike length Fruiting efficiency Seed Width Seed Length Seed Area cm from the tip of the spike to the end of the awn g average weight of 3 reps of 100 kernels cm from the last node to the base of cm from the auricle of the flag leaf to the base of cm from base to tip of Max=17.0; Mean=9.8 Min=22.5; Max=73.0; Mean=51.1 Min=2.8; Max=34.5; Mean=13.4 Min=3.3; Max=14.3; Mean=8.3 Min=5; Max=44; Mean=19.8 Min=6; Max=45; Mean=25.0 Grain/spike weight ratio mm Marvin at JHI, mm Marvin at JHI, mm2 Marvin at JHI, Min=2.2; Max=4.3; Mean=3.6 Min=6.6; Max=18.9; Mean=9.9 Min=14.6; Max=42.5; Mean=26.0 Meteo data Available Available Available Available Available Sowing date 31/03/ /04/ /03/ /03/ /03/2015 Notes Very late sowing, uneven heading Severe lodging, not possible to harvest Irrigated Rainfed Seeds Not harvested 10 ears harvested Not harvested Stored Stored 7 / 10

8 mandatory traits additional traits Wheat Winter WW1 WW2 WW3 WW4 WW5 Variable Name Description INRAFRA KWSUK MTAHUN TUR TUR Plant Establishment Visual score (03) Winter survival Visual score (09) Heading date Plant height Awn length Awn roughness (1000)Grain weight Peduncle length Nr. Of days from sowing to Zadoks DGS 53 cm from soil to the tip of cm from the tip of the spike to the end of the awn 1: Smooth; 2: smooth base/rough tip; 3: rough g average weight of 3 reps of 100 kernels cm from the last node to the base of Lodging intensity Visual score (19) Spike length Fruiting efficiency cm from base to tip of Max=2; Mean=1.3 Max=4; Mean=1.3 Min=142; Max=186; Mean=159.0 Min=48; Max=174; Mean=103.3 Max=13.7; Mean=3.8 Mean=2.96 Min=22.4; Max=73.6; Mean=43.8 Min=195; Max=236; Mean=215.5 Min=40; Max=185; Mean=108.3 Presence vs. Absence of awns Mean=2.7 Max=9; Mean=0.6 Min=197; Max=237; Mean=212.7 Min=41; Max=149; Mean=95.8 Max=12.5; Mean=3.1 Mean=1.9 Max=2; Mean=1.3 Min=187; Max=210; Mean=197.1 Min=22; Max=144.5; Mean=90.8 Min=177; Max=200; Mean=193.5 Min=25.5; Max=130; Mean=86.4 Min=15.1; Max=69.1; Mean=39.8 Max=9; Mean=1.8 Grain/spike weight ratio Yellow rust Visual score (0100%) Carbon isotope discrimination Carbon content Percentage (%) Leaf rust Visual score (19) Stripe rust Visual score (19) Powdery mildew Visual score (19) Meteo data Min=31.2; Max=25.9; Mean=29.6 Min=30.7; Max=55.5; Mean=43.9 Mean=1.0 Max=9; Mean=1.5 Max=5; Mean=1.04 Available, along with soil information Min=5.0; Max=50.0; Mean=26.4 Min=7.0; Max=60.0; Mean=28.3 Max=100; Mean=59.0 Min=3.9; Max=18.0; Mean=10.7 Min=0.51; Max=0.97; Mean=0.71 Only temperature Available Available Available Sowing date 510/12/ /10/ /10/ /11/ /11/2014 Notes Seeds Stored + sent to INRA Different sowing plan Not harvested Irrigated Rainfed Stored + sent to UZH and DLO Stored Stored 8 / 10

9 mandatory traits Wheat Spring WS2 WS3 WS4 Variable Name Description MTAHUN TUR TUR Plant Establishment Heading date Plant height Awn length Visual score (03) Nr. Of days from sowing to Zadoks DGS 53 cm from soil to the tip of the spike, awn excluded cm from the tip of to the end of the awn Mean=2.5 Min=74; Max=112; Mean=88.4; 40% of plots did not flower Min=35; Max=118; Mean=81.7 Max=1; Mean=0.9 Max=1; Mean=0.9 Min=20; Max=124; Mean=57.4 Min=24; Max=120; Mean=69.5 (1000)Grain weight Peduncle length g average weight of 3 reps of 100 kernels cm from the last node to the base of Min=3; Max=64; Mean=23.2 Min=5; Max=56; Mean=27.5 Meteo data Available Available Available Sowing date 11/03/ /03/ /03/2015 Notes Irrigated Rainfed Seeds Not harvested Stored Stored 9 / 10

10 1.4. Conclusions The phenotypic data acquired highlight the wide genetic diversity present in the WHEALBI germplasm, suggesting that different genetic factors are responsible for the adaptation of each genotype to specific environmental conditions. These data will be analyzed together with the molecular data generated by exome capture and resequencing in WP2 to identify the genes responsible of specific adaptive traits. The garden experiments have also served as multiplication of all accessions in order to produce enough seeds for the phenotyping activities organized in the next three years of the project. 10 / 10